Exosomal Cripto-1 Serves as a Potential Biomarker for Perihilar Cholangiocarcinoma

Abstract

Perihilar cholangiocarcinoma (PHCCA) has a poor prognosis, mainly due to diagnosis at an advanced stage. Cripto-1 functions as an oncogene and is highly expressed in several human cancers, however, its clinical application in PHCCA is poorly understood. Herein, we identified that Cripto-1 was released by PHCCA cells via exosomes in vitro and in vivo. Furthermore, an ELISA method was developed to detect exosomal Cripto-1 in the serum of 115 PHCCA patients, 47 cholangitis patients and 65 healthy controls, and it was found that exosomal Cripto-1 was increased in PHCCA patients and associated with metastasis. Compared with traditional serum tumor markers, CA19-9 and CEA, exosomal Cripto-1 demonstrated a larger area under ROC curve for PHCCA diagnosis. The cutoff value of exosomal Cripto-1 was 0.82, achieving a sensitivity of 79.1% and a specificity of 87.5%. As expected, exosomal Cripto-1 levels in immunohistochemically Cripto-1-high cases were significantly elevated compared to in Cripto-1-low cases. When measured 1-week postoperatively, Cripto-1 levels decreased on average from 1.25(0.96-3.26) to 0.85(0.62-1.82). Immunohistochemistry analysis showed Cripto-1 expression was negatively correlated with E-cadherin and was an independent prognostic biomarker for poor survival in PHCCA patients. In conclusion, exosomal Cripto-1 in sera can reflect its expression in the tissue of PHCAA patients and has the potential be a non-invasive biomarker for diagnosis and prognosis of PHCCA.

A Novel Localization in Human Large Extracellular Vesicles for the EGF-CFC Founder Member CRIPTO and Its Biological and Therapeutic Implications

“…In agreement, several studies have reported the presence of various GPI-anchored proteins also in exosomes [ 60 ]. In particular, very recently CRIPTO has been found on the surface on sEVs released by another type of cancer cell: the perihilar cholangiocarcinoma (PHCCA) cells [ 20 ]. Our analysis showing CRIPTO also in sEVs and lEVs derived from teratocarcinoma cells, coupled with the previous finding of Hu and coauthors (2021) [ 20 ], led us to hypothesize a more general mechanism that could involve many types of cancer.…”

“…In particular, very recently CRIPTO has been found on the surface on sEVs released by another type of cancer cell: the perihilar cholangiocarcinoma (PHCCA) cells [ 20 ]. Our analysis showing CRIPTO also in sEVs and lEVs derived from teratocarcinoma cells, coupled with the previous finding of Hu and coauthors (2021) [ 20 ], led us to hypothesize a more general mechanism that could involve many types of cancer. Noteworthy, CRIPTO might be an EV marker present on the surface of vesicles released by different types of cancer cells.…”

“…Therefore, Cripto can act both in cis as a membrane bound protein as well as in trans as a soluble factor. Very recently, Hu and coauthors reported also the presence of CRIPTO on the membrane of the sEVs released by perihilar cholangiocarcinoma (PHCCA) cells [ 20 ].…”

Extracellular vesicle isolation, purification and evaluation in cancer diagnosis

“…Cripto-1, for instance, is expressed at high levels in several number of human cancers (Ref. 91). However, its evaluation can be an indicative of poor prognosis, and it can be assessed as a promising marker for early cancer detection.…”

“…Exosomal Cripto-1 can be assessed in serum of patients with perihilar cholangiocarcinoma, and it can be considered as a potential biomarker (Ref. 91). Exosomal HOXD-AS1 is increased in serum of patients with prostatic cancer, and is regarded as a promising biomarker for diagnosis and treatment of the metastatic cancer.…”